TY - JOUR
T1 - Specific combinations of presenilins and Aph1s affect the substrate specificity and activity of γ-secretase
AU - Yonemura, Yoji
AU - Futai, Eugene
AU - Yagishita, Sosuke
AU - Kaether, Christoph
AU - Ishiura, Shoichi
N1 - Funding Information:
Y.Y. was supported by Japan Society for the Promotion of Science . This work was supported in part by an Intramural Research Grant ( MHLW; 26-8 ) for Neurological and Psychiatric Disorders of NCNP, a research grant for Comprehensive Research on Disability Health and Welfare from the Ministry of Health, Labour and Welfare (MHLW; H26-Sinkeikinn-ippan-004) and a Grant-in-Aid from the MHLW of Japan (to S.I.).
Funding Information:
To express six different γ-secretases and Gal4 fusion substrates in yeast, PS (PS1 or PS2), NCT, Aph1-HA (Aph1aL-HA or Aph1aS-HA or Aph1b-HA), Flag-Pen2 and C55-Gal4 or Notch-Gal4 were cloned into the pBEVY, as described previously [11] . An Aph1b clone (GNP Clone IRAL037F15) was provided by the RIKEN Bio Resource Center, which participates with the National Bio Resource Project of the Ministry of Education, Culture, Sports, Science and Technology of Japan [13–16] .
Publisher Copyright:
© 2016 The Authors
PY - 2016/9/30
Y1 - 2016/9/30
N2 - The γ-secretase complex comprises presenilin (PS), nicastrin (NCT), anterior pharynx-defective 1 (Aph1), and presenilin enhancer 2 (Pen2). PS has two homologues, PS1 and PS2. Aph1 has two isoforms, Aph1a and Aph1b, with the former existing as two splice variants Aph1aL and Aph1aS. Each complex consists of one subunit each, resulting in six different γ-secretases. To better understand the functional differences among the γ-secretases, we reconstituted them using a yeast system and compared Notch1-cleavage and amyloid precursor protein (APP)-cleavage activities. Intriguingly, PS2/Aph1b had a clear substrate specificity: APP-Gal4, but not Notch-Gal4, was cleaved. In HEK cell lines expressing defined γ-secretase subunits, we showed that PS1/Aph1b, PS2/Aph1aL, PS2/Aph1aS and PS2/Aph1b γ-secretase produced amyloid β peptide (Aβ) with a higher Aβ42+Aβ43-to-Aβ40 (Aβ42(43)/Aβ40) ratio than the other γ-secretases. In addition, PS2/Aph1aS γ-secretase produced less Notch intracellular domain (NICD) than did the other 5 γ-secretases. Considering that the Aβ42(43)/Aβ40 ratio is relevant in the pathogenesis of Alzheimer's disease (AD), and that inhibition of Notch cleavage causes severe side effect, these results suggest that the PS2/Aph1aS γ-secretase complex is a potential therapeutic target in AD.
AB - The γ-secretase complex comprises presenilin (PS), nicastrin (NCT), anterior pharynx-defective 1 (Aph1), and presenilin enhancer 2 (Pen2). PS has two homologues, PS1 and PS2. Aph1 has two isoforms, Aph1a and Aph1b, with the former existing as two splice variants Aph1aL and Aph1aS. Each complex consists of one subunit each, resulting in six different γ-secretases. To better understand the functional differences among the γ-secretases, we reconstituted them using a yeast system and compared Notch1-cleavage and amyloid precursor protein (APP)-cleavage activities. Intriguingly, PS2/Aph1b had a clear substrate specificity: APP-Gal4, but not Notch-Gal4, was cleaved. In HEK cell lines expressing defined γ-secretase subunits, we showed that PS1/Aph1b, PS2/Aph1aL, PS2/Aph1aS and PS2/Aph1b γ-secretase produced amyloid β peptide (Aβ) with a higher Aβ42+Aβ43-to-Aβ40 (Aβ42(43)/Aβ40) ratio than the other γ-secretases. In addition, PS2/Aph1aS γ-secretase produced less Notch intracellular domain (NICD) than did the other 5 γ-secretases. Considering that the Aβ42(43)/Aβ40 ratio is relevant in the pathogenesis of Alzheimer's disease (AD), and that inhibition of Notch cleavage causes severe side effect, these results suggest that the PS2/Aph1aS γ-secretase complex is a potential therapeutic target in AD.
KW - Alzheimer's disease
KW - Aβ
KW - Notch
KW - γ-secretase
UR - http://www.scopus.com/inward/record.url?scp=84992013304&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84992013304&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2016.09.018
DO - 10.1016/j.bbrc.2016.09.018
M3 - Article
C2 - 27608597
AN - SCOPUS:84992013304
SN - 0006-291X
VL - 478
SP - 1751
EP - 1757
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 4
ER -